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Soluble TNF-α Receptor I Encoded on Plasmid Vector and Its Application in Experimental Gene Therapy of Radiation-Induced Lung Fibrosis

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Archivum Immunologiae et Therapiae Experimentalis Aims and scope

Abstract

Post-radiation inflammatory reaction leads to an irreversible pulmonary fibrosis which may cause lethal respiratory insufficiency. Pathological inflammatory and fibrotic changes might be attenuated by inhibiting tumour necrosis factor (TNF)-α activity using TNF-α soluble receptors. Thus, an experimental antifibrotic gene therapy with the plasmid vector encoding a mouse soluble receptor I for TNF-α (psTNFR-I) was assessed. Soluble TNFR-I encoding gene was cloned into pcDNA3.1 plasmid. The ability of psTNFR-I expressing vector to transfect cells, and its biological activity in vitro and in vivo were examined by PCR, RT-PCR, MTT assay and ELISA. The C57Bl/6J mice received single intramuscular injection of psTNFR-I, conjugated with polyetylenimine (PEI) 25 kDa, equally divided to both hind legs, 3 days before irradiation (20 Gy, Co60), and either a single injection or ten injections once a week after irradiation. The data proved the effectiveness of psTNFR-I product to neutralise TNF-α activity in vitro. The in vivo plasmid incorporation and maintenance was confirmed. Measurements of plasma soluble TNFR-I levels showed that the in vivo gene transfer was effective. PEI was found to enhance transfection efficiency in vivo. The psTNFR-I/PEI complexes caused no toxicity in the transfected mice. C57Bl/6J mice that received prolonged psTNFR-I/PEI injections developed lethal fibrotic syndrome and died 8 weeks later than the mice treated with a double plasmid injection and the control mice treated with a control plasmid. Sequential administration of soluble TNFR-I by a nonviral, intramuscular gene transduction in the early and late post-radiation inflammatory phase prolonged survival of irradiated mice and attenuated the symptoms of lung fibrosis. The psTNFR-I gene transduction may provide a safe and simple method to partially neutralise TNF-α activity and prevent radiation-induced lung injury.

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Acknowledgments

This work was supported by Polish Ministry of Science and Higher Education grant N401 077 32/1925. The authors thank Dr. Dariusz Kowalczyk for valuable contribution in this project and Dr. Magdalena Chechlińska, for helpful comments on the manuscript.

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Authors and Affiliations

  1. Cell Biology Department, Maria Skłodowska-Curie Memorial Institute of Oncology and Cancer Centre, W. K. Roentgena 5, 02-781, Warsaw, Poland

    Małgorzata Przybyszewska, Joanna Miłoszewska, Sylwia Rzońca, Halina Trembacz, Agnieszka Kotlarz, Paweł Swoboda & Maciej Małecki

  2. Department of Genetic and Animals Breeding, Maria Skłodowska-Curie Memorial Institute of Oncology and Cancer Centre, Warsaw, Poland

    Kazimiera Pyśniak

  3. Medical Physics Department, Maria Skłodowska-Curie Memorial Institute of Oncology and Cancer Centre, Warsaw, Poland

    Marta Zalewska

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  1. Małgorzata Przybyszewska
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Correspondence to Małgorzata Przybyszewska.

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Przybyszewska, M., Miłoszewska, J., Rzońca, S. et al. Soluble TNF-α Receptor I Encoded on Plasmid Vector and Its Application in Experimental Gene Therapy of Radiation-Induced Lung Fibrosis. Arch. Immunol. Ther. Exp. 59, 315–326 (2011). https://doi.org/10.1007/s00005-011-0133-2

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